CN115304616B - Gastrodia elata active compound and preparation method and application thereof - Google Patents
Gastrodia elata active compound and preparation method and application thereof Download PDFInfo
- Publication number
- CN115304616B CN115304616B CN202211104188.5A CN202211104188A CN115304616B CN 115304616 B CN115304616 B CN 115304616B CN 202211104188 A CN202211104188 A CN 202211104188A CN 115304616 B CN115304616 B CN 115304616B
- Authority
- CN
- China
- Prior art keywords
- gel column
- component
- chloroform
- methanol
- silica gel
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 241000305491 Gastrodia elata Species 0.000 title claims abstract description 65
- 150000001875 compounds Chemical class 0.000 title claims abstract description 65
- 238000002360 preparation method Methods 0.000 title claims abstract description 29
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 53
- 238000000926 separation method Methods 0.000 claims abstract description 23
- 238000000605 extraction Methods 0.000 claims abstract description 21
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 162
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 82
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 78
- 238000013375 chromatographic separation Methods 0.000 claims description 72
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 62
- 239000000741 silica gel Substances 0.000 claims description 62
- 229910002027 silica gel Inorganic materials 0.000 claims description 62
- 238000010828 elution Methods 0.000 claims description 61
- 239000002904 solvent Substances 0.000 claims description 61
- 239000000499 gel Substances 0.000 claims description 55
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 54
- 239000003480 eluent Substances 0.000 claims description 54
- 238000010898 silica gel chromatography Methods 0.000 claims description 37
- WORJEOGGNQDSOE-UHFFFAOYSA-N chloroform;methanol Chemical compound OC.ClC(Cl)Cl WORJEOGGNQDSOE-UHFFFAOYSA-N 0.000 claims description 36
- 238000004440 column chromatography Methods 0.000 claims description 36
- 239000012046 mixed solvent Substances 0.000 claims description 24
- 239000003208 petroleum Substances 0.000 claims description 24
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 20
- OAIVIYSBZFEOIU-UHFFFAOYSA-N chloroform;propan-2-one Chemical compound CC(C)=O.ClC(Cl)Cl OAIVIYSBZFEOIU-UHFFFAOYSA-N 0.000 claims description 20
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 15
- HWJHWSBFPPPIPD-UHFFFAOYSA-N ethoxyethane;propan-2-one Chemical compound CC(C)=O.CCOCC HWJHWSBFPPPIPD-UHFFFAOYSA-N 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 238000004042 decolorization Methods 0.000 claims description 9
- 238000000034 method Methods 0.000 claims description 9
- NBEMQPLNBYYUAZ-UHFFFAOYSA-N ethyl acetate;propan-2-one Chemical compound CC(C)=O.CCOC(C)=O NBEMQPLNBYYUAZ-UHFFFAOYSA-N 0.000 claims description 8
- 239000000284 extract Substances 0.000 claims description 7
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 claims description 6
- PBCJIPOGFJYBJE-UHFFFAOYSA-N acetonitrile;hydrate Chemical compound O.CC#N PBCJIPOGFJYBJE-UHFFFAOYSA-N 0.000 claims description 5
- 238000002953 preparative HPLC Methods 0.000 claims description 5
- 239000000469 ethanolic extract Substances 0.000 claims description 4
- 229940122041 Cholinesterase inhibitor Drugs 0.000 claims description 3
- 239000000544 cholinesterase inhibitor Substances 0.000 claims description 3
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 3
- 230000001404 mediated effect Effects 0.000 claims description 3
- 238000010992 reflux Methods 0.000 claims description 3
- 208000024827 Alzheimer disease Diseases 0.000 claims 1
- 206010039966 Senile dementia Diseases 0.000 claims 1
- 239000003814 drug Substances 0.000 abstract description 10
- 230000000694 effects Effects 0.000 abstract description 9
- 239000002994 raw material Substances 0.000 abstract description 7
- 229940079593 drug Drugs 0.000 abstract description 6
- 230000005764 inhibitory process Effects 0.000 abstract description 4
- 102000003914 Cholinesterases Human genes 0.000 abstract description 2
- 108090000322 Cholinesterases Proteins 0.000 abstract description 2
- 102000004190 Enzymes Human genes 0.000 abstract description 2
- 108090000790 Enzymes Proteins 0.000 abstract description 2
- 229940048961 cholinesterase Drugs 0.000 abstract description 2
- 229940088598 enzyme Drugs 0.000 abstract description 2
- 241000411851 herbal medicine Species 0.000 abstract description 2
- 238000009776 industrial production Methods 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 20
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 16
- XELZGAJCZANUQH-UHFFFAOYSA-N methyl 1-acetylthieno[3,2-c]pyrazole-5-carboxylate Chemical compound CC(=O)N1N=CC2=C1C=C(C(=O)OC)S2 XELZGAJCZANUQH-UHFFFAOYSA-N 0.000 description 12
- 239000002245 particle Substances 0.000 description 9
- YLJREFDVOIBQDA-UHFFFAOYSA-N tacrine Chemical compound C1=CC=C2C(N)=C(CCCC3)C3=NC2=C1 YLJREFDVOIBQDA-UHFFFAOYSA-N 0.000 description 7
- 229960001685 tacrine Drugs 0.000 description 7
- 102100033639 Acetylcholinesterase Human genes 0.000 description 6
- 108010022752 Acetylcholinesterase Proteins 0.000 description 6
- 238000002835 absorbance Methods 0.000 description 5
- 239000007864 aqueous solution Substances 0.000 description 5
- OKKJLVBELUTLKV-VMNATFBRSA-N methanol-d1 Chemical compound [2H]OC OKKJLVBELUTLKV-VMNATFBRSA-N 0.000 description 5
- 229940022698 acetylcholinesterase Drugs 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- SRCZQMGIVIYBBJ-UHFFFAOYSA-N ethoxyethane;ethyl acetate Chemical compound CCOCC.CCOC(C)=O SRCZQMGIVIYBBJ-UHFFFAOYSA-N 0.000 description 4
- 238000011894 semi-preparative HPLC Methods 0.000 description 4
- OKKJLVBELUTLKV-MZCSYVLQSA-N Deuterated methanol Chemical compound [2H]OC([2H])([2H])[2H] OKKJLVBELUTLKV-MZCSYVLQSA-N 0.000 description 3
- 241000305492 Gastrodia Species 0.000 description 3
- 229930191978 Gibberellin Natural products 0.000 description 3
- IXORZMNAPKEEDV-UHFFFAOYSA-N gibberellic acid GA3 Natural products OC(=O)C1C2(C3)CC(=C)C3(O)CCC2C2(C=CC3O)C1C3(C)C(=O)O2 IXORZMNAPKEEDV-UHFFFAOYSA-N 0.000 description 3
- 239000003448 gibberellin Substances 0.000 description 3
- 238000010829 isocratic elution Methods 0.000 description 3
- 239000008363 phosphate buffer Substances 0.000 description 3
- 230000002792 vascular Effects 0.000 description 3
- KIUMMUBSPKGMOY-UHFFFAOYSA-N 3,3'-Dithiobis(6-nitrobenzoic acid) Chemical compound C1=C([N+]([O-])=O)C(C(=O)O)=CC(SSC=2C=C(C(=CC=2)[N+]([O-])=O)C(O)=O)=C1 KIUMMUBSPKGMOY-UHFFFAOYSA-N 0.000 description 2
- PUQSUZTXKPLAPR-KSSYENDESA-N 4-(beta-D-Glucopyranosyloxy) benzyl alcohol Natural products O([C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1)c1ccc(CO)cc1 PUQSUZTXKPLAPR-KSSYENDESA-N 0.000 description 2
- HDHRTQZSBFUBMJ-UHFFFAOYSA-N Artonin E Natural products O1C2=C3C=CC(C)(C)OC3=CC(O)=C2C(=O)C(CC=C(C)C)=C1C1=CC(O)=C(O)C=C1O HDHRTQZSBFUBMJ-UHFFFAOYSA-N 0.000 description 2
- 206010010904 Convulsion Diseases 0.000 description 2
- PUQSUZTXKPLAPR-UJPOAAIJSA-N Gastrodin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=C(CO)C=C1 PUQSUZTXKPLAPR-UJPOAAIJSA-N 0.000 description 2
- GOBAQYCCUYZMJY-DKEHAURLSA-N Mulberrofuran K Natural products Oc1c2[C@H]3[C@@H]4[C@](c5c6OC(C)(C)C=Cc6c(O)cc5)(Oc2cc(-c2oc5c(c2)ccc(O)c5)c1)Oc1c([C@H]4CC(C)=C3)ccc(O)c1 GOBAQYCCUYZMJY-DKEHAURLSA-N 0.000 description 2
- 230000036461 convulsion Effects 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- 229930193974 gastrodin Natural products 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- PUQSUZTXKPLAPR-NZEXEKPDSA-N helicidol Natural products O([C@H]1[C@H](O)[C@H](O)[C@@H](O)[C@@H](CO)O1)c1ccc(CO)cc1 PUQSUZTXKPLAPR-NZEXEKPDSA-N 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- XFFOMNJIDRDDLQ-UHFFFAOYSA-N morusin Chemical compound O1C2=C3C=CC(C)(C)OC3=CC(O)=C2C(=O)C(CC=C(C)C)=C1C1=CC=C(O)C=C1O XFFOMNJIDRDDLQ-UHFFFAOYSA-N 0.000 description 2
- WUBUWBUVAKMGCO-UHFFFAOYSA-N morusin Natural products CC(=CCC1=C(Cc2c3C=CC(C)(C)Oc3cc(O)c2C1=O)c4ccc(O)cc4O)C WUBUWBUVAKMGCO-UHFFFAOYSA-N 0.000 description 2
- 229930186407 mulberrofuran Natural products 0.000 description 2
- BVJSUAQZOZWCKN-UHFFFAOYSA-N p-hydroxybenzyl alcohol Chemical compound OCC1=CC=C(O)C=C1 BVJSUAQZOZWCKN-UHFFFAOYSA-N 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000012224 working solution Substances 0.000 description 2
- CSCPPACGZOOCGX-MICDWDOJSA-N 1-deuteriopropan-2-one Chemical compound [2H]CC(C)=O CSCPPACGZOOCGX-MICDWDOJSA-N 0.000 description 1
- LDHMAVIPBRSVRG-UHFFFAOYSA-O 1-methylnicotinamide Chemical compound C[N+]1=CC=CC(C(N)=O)=C1 LDHMAVIPBRSVRG-UHFFFAOYSA-O 0.000 description 1
- SZMVXHRECFRCKQ-UHFFFAOYSA-M 2-ethanethioyloxyethyl(trimethyl)azanium;iodide Chemical compound [I-].CC(=S)OCC[N+](C)(C)C SZMVXHRECFRCKQ-UHFFFAOYSA-M 0.000 description 1
- 241001083548 Anemone Species 0.000 description 1
- 206010019233 Headaches Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- 206010033799 Paralysis Diseases 0.000 description 1
- 208000005392 Spasm Diseases 0.000 description 1
- 241000001727 Tropicoporus linteus Species 0.000 description 1
- CSCPPACGZOOCGX-WFGJKAKNSA-N acetone d6 Chemical compound [2H]C([2H])([2H])C(=O)C([2H])([2H])[2H] CSCPPACGZOOCGX-WFGJKAKNSA-N 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 230000001914 calming effect Effects 0.000 description 1
- 230000002490 cerebral effect Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 208000002173 dizziness Diseases 0.000 description 1
- 206010015037 epilepsy Diseases 0.000 description 1
- 238000003810 ethyl acetate extraction Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 231100000869 headache Toxicity 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- -1 isopentenyl flavonoid compounds Chemical class 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 125000000896 monocarboxylic acid group Chemical group 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 231100000862 numbness Toxicity 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000002936 tranquilizing effect Effects 0.000 description 1
- CTGNYPVJSIRPLG-UHFFFAOYSA-N trimethyl(2-sulfanylethyl)azanium;iodide Chemical compound [I-].C[N+](C)(C)CCS CTGNYPVJSIRPLG-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
- C07D493/02—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
- C07D493/06—Peri-condensed systems
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/898—Orchidaceae (Orchid family)
- A61K36/8988—Gastrodia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C45/00—Preparation of compounds having >C = O groups bound only to carbon or hydrogen atoms; Preparation of chelates of such compounds
- C07C45/78—Separation; Purification; Stabilisation; Use of additives
- C07C45/79—Separation; Purification; Stabilisation; Use of additives by solid-liquid treatment; by chemisorption
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C49/00—Ketones; Ketenes; Dimeric ketenes; Ketonic chelates
- C07C49/587—Unsaturated compounds containing a keto groups being part of a ring
- C07C49/753—Unsaturated compounds containing a keto groups being part of a ring containing ether groups, groups, groups, or groups
- C07C49/755—Unsaturated compounds containing a keto groups being part of a ring containing ether groups, groups, groups, or groups a keto group being part of a condensed ring system with two or three rings, at least one ring being a six-membered aromatic ring
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
- C07D307/77—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D307/78—Benzo [b] furans; Hydrogenated benzo [b] furans
- C07D307/79—Benzo [b] furans; Hydrogenated benzo [b] furans with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to carbon atoms of the hetero ring
- C07D307/80—Radicals substituted by oxygen atoms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
- C07D493/02—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
- C07D493/04—Ortho-condensed systems
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C2603/00—Systems containing at least three condensed rings
- C07C2603/02—Ortho- or ortho- and peri-condensed systems
- C07C2603/04—Ortho- or ortho- and peri-condensed systems containing three rings
- C07C2603/06—Ortho- or ortho- and peri-condensed systems containing three rings containing at least one ring with less than six ring members
- C07C2603/10—Ortho- or ortho- and peri-condensed systems containing three rings containing at least one ring with less than six ring members containing five-membered rings
- C07C2603/12—Ortho- or ortho- and peri-condensed systems containing three rings containing at least one ring with less than six ring members containing five-membered rings only one five-membered ring
- C07C2603/16—Benz[e]indenes; Hydrogenated benz[e]indenes
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Gastroenterology & Hepatology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Epidemiology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention provides a gastrodia elata active compound and a preparation method and application thereof, and relates to the technical field of biological medicines. The gastrodia elata active compound provided by the invention has the structure shown in the formulas I-III and V, has a good enzyme activity effect, particularly has a good inhibition effect on cholinesterase, and has a good application prospect in the aspect of preparing medicines. The gastrodia elata active compound is obtained by alcohol extraction and separation of the traditional Chinese medicine gastrodia elata serving as a raw material, is derived from Chinese herbal medicines, has high safety and effectiveness, and greatly expands the pharmaceutical economic value of gastrodia elata; the preparation method is simple to operate, low in cost and low in cost, and raw materials are cheap and easy to obtain, so that the preparation method is suitable for industrial production; in addition, the preparation method provided by the invention can avoid using a large amount of organic raw materials, and is environment-friendly.
Description
Technical Field
The invention relates to the technical field of biological medicines, in particular to a gastrodia elata active compound and a preparation method and application thereof.
Background
The traditional Chinese medicine gastrodia elata (called red arrow, fixed anemone, etc.) is a dry tuber of Gastrodia elata Bl. Belonging to Gastrodia genus (Gastrodia R.Br.) of Orhidaceae family, and has effects of calming endogenous wind, relieving spasm, suppressing liver yang, dispelling pathogenic wind, and dredging collaterals. Is mainly used for treating liver wind internal movement, convulsion, epilepsy, convulsion, dizziness, headache, limb numbness, hand and foot paralysis, rheumatalgia and the like, and is a traditional rare traditional Chinese medicine. Currently, the main chemical components of gastrodia genus reported in literature are more than eighty compounds including phenolic components such as gastrodin and p-hydroxy benzyl alcohol, derivatives thereof, polysaccharides and polypeptides. The main chemical components have various biological activities, such as tranquilizing and allaying excitement, reducing peripheral vascular, cerebral vascular and coronary vascular resistance, lowering blood pressure, slowing heart rate, easing pain and resisting inflammation, and the gastrodia elata polysaccharide has immunological activity.
At present, only gastrodin (gibberellin) is reported in gastrodia elata, and other separation and identification of the gibberellin and the phellinus linteus are not reported, so that research on new active ingredients in gastrodia elata has important significance for expanding the application range of gastrodia elata.
Disclosure of Invention
In view of the above, the invention aims to provide a gastrodia elata active compound, a preparation method and application thereof.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a gastrodia elata active compound, which has a structure shown in any one of formulas I-III:
the invention provides a preparation method of the gastrodia elata active compound, which comprises the following steps:
(1) Extracting gastrodia elata with alcohol to obtain an alcohol extract; extracting the ethanol extract with ethyl acetate, and concentrating to obtain an ethyl acetate phase;
(2) Subjecting the ethyl acetate phase to first silica gel column chromatography separation to obtain a component Fr.8; the eluent adopted by the first silica gel column chromatographic separation is petroleum ether-acetone solvent, the volume ratio of petroleum ether to acetone in the petroleum ether-acetone solvent is 100:1-0:1, and the elution mode is gradient elution;
(3) Subjecting the component Fr.8 to a second silica gel column chromatography to obtain a component Fr.8.6 and a component Fr.8.8; the eluent adopted by the second silica gel column chromatographic separation is chloroform-acetone solvent, the volume ratio of chloroform to acetone in the chloroform-acetone solvent is 15:1-0:1, and the elution mode is gradient elution;
(4) Subjecting the component Fr.8.6 to third silica gel column chromatography to obtain a component Fr.8.6.6; the eluent adopted by the third silica gel column chromatographic separation is chloroform-acetone solvent, the volume ratio of chloroform to acetone in the chloroform-acetone solvent is 5:1-2:1, and the elution mode is gradient elution;
sequentially performing first SephadexLH-20 gel column chromatography, fourth silica gel column chromatography, second SephadexLH-20 gel column chromatography and semi-preparative high performance liquid chromatography on the component Fr.8.6.6 to obtain rhizoma Gastrodiae active compounds with structures shown in formula I and formula II; the eluent adopted by the first SephadexLH-20 gel column chromatography is a chloroform-methanol mixed solvent, and the volume ratio of chloroform to methanol in the chloroform-methanol mixed solvent is 1:1; the eluent adopted by the fourth silica gel column chromatographic separation is ethyl acetate-acetone solvent, the volume ratio of ethyl acetate to acetone in the ethyl acetate-acetone solvent is 1:2-1:3, and the elution mode is gradient elution; the eluent adopted by the chromatographic separation of the second SephadexLH-20 gel column is methanol; the mobile phase adopted by the semi-preparation high performance liquid chromatography separation is acetonitrile water solution with the acetonitrile volume fraction of 65%;
(5) Performing MCI (MCI-mediated color removal) on the component Fr.8.8 to obtain a component Fr.8.8.1;
sequentially performing fifth silica gel column chromatographic separation, third Sephadex LH-20 gel column chromatographic separation and fourth Sephadex LH-20 gel column chromatographic separation on the component Fr.8.8.1 to obtain a gastrodia elata active compound with a structure shown in a formula III; the eluent adopted by the fifth silica gel column chromatographic separation is chloroform-methanol solvent, the volume ratio of chloroform to methanol in the chloroform-methanol solvent is 6:1-0:1, and the elution mode is gradient elution; the eluent adopted by the third SephadexLH-20 gel column chromatography is a chloroform-methanol mixed solvent, and the volume ratio of chloroform to methanol in the chloroform-methanol mixed solvent is 1:1; the eluent adopted by the chromatographic separation of the fourth SephadexLH-20 gel column is methanol;
the step (4) and the step (5) have no time sequence.
Preferably, the alcohol extraction is performed with an aqueous alcohol solution; the alcohol in the alcohol-water solution comprises ethanol and/or methanol; the volume fraction of alcohol in the aqueous alcohol solution is 95%.
Preferably, the ratio of the mass of the gastrodia elata to the volume of the alcohol-water solution is 1kg: 2-4L.
Preferably, the alcohol extraction is reflux alcohol extraction; the times of the alcohol extraction are 2-3 times, and the time of the single alcohol extraction is 1-3 hours.
Preferably, the eluent used for MCI color removal is a water-methanol solvent, and the volume ratio of water to methanol in the water-methanol solvent is 7:3-0:1.
The invention provides the gastrodia elata active compound disclosed by the technical scheme, the gastrodia elata active compound prepared by the preparation method disclosed by the technical scheme, or the application of any one of the gastrodia elata active compounds with structures shown in formulas IV-VII in preparation of medicines;
preferably, the use is as cholinesterase inhibitor.
The invention provides a gastrodia elata active compound, which has a structure shown in any one of formulas I-III. The gastrodia elata active compound provided by the invention has a good enzyme activity effect, particularly has a good inhibition effect on cholinesterase, and has a good application prospect in the aspect of preparing medicines. In addition, the gastrodia elata active compound provided by the invention can be applied to medical instrument products.
The invention provides a preparation method of the gastrodia elata active compound. The traditional Chinese medicine gastrodia elata is taken as a raw material for alcohol extraction and separation to obtain the gastrodia elata active compound, and the gastrodia elata active compound is derived from Chinese herbal medicines, so that the safety and the effectiveness are high, and the pharmaceutical economic value of the gastrodia elata is greatly expanded; the preparation method is simple to operate, low in cost and low in cost, and raw materials are cheap and easy to obtain, so that the preparation method is suitable for industrial production; in addition, the preparation method provided by the invention can avoid using a large amount of organic raw materials, and is environment-friendly.
Detailed Description
The invention provides a gastrodia elata active compound, which has a structure shown in any one of formulas I-III:
the invention provides a preparation method of the gastrodia elata active compound, which comprises the following steps:
(1) Extracting gastrodia elata with alcohol to obtain an alcohol extract; extracting the ethanol extract with ethyl acetate, and concentrating to obtain an ethyl acetate phase;
(2) Subjecting the ethyl acetate phase to first silica gel column chromatography separation to obtain a component Fr.8; the eluent adopted by the first silica gel column chromatographic separation is petroleum ether-acetone solvent, the volume ratio of petroleum ether to acetone in the petroleum ether-acetone solvent is 100:1-0:1, and the elution mode is gradient elution;
(3) Subjecting the component Fr.8 to a second silica gel column chromatography to obtain a component Fr.8.6 and a component Fr.8.8; the eluent adopted by the second silica gel column chromatographic separation is chloroform-acetone solvent, the volume ratio of chloroform to acetone in the chloroform-acetone solvent is 15:1-0:1, and the elution mode is gradient elution;
(4) Subjecting the component Fr.8.6 to third silica gel column chromatography to obtain a component Fr.8.6.6; the eluent adopted by the third silica gel column chromatographic separation is chloroform-acetone solvent, the volume ratio of chloroform to acetone in the chloroform-acetone solvent is 5:1-2:1, and the elution mode is gradient elution;
sequentially performing first SephadexLH-20 gel column chromatography, fourth silica gel column chromatography, second SephadexLH-20 gel column chromatography and semi-preparative high performance liquid chromatography on the component Fr.8.6.6 to obtain rhizoma Gastrodiae active compounds with structures shown in formula I and formula II; the eluent adopted by the first SephadexLH-20 gel column chromatography is a chloroform-methanol mixed solvent, and the volume ratio of chloroform to methanol in the chloroform-methanol mixed solvent is 1:1; the eluent adopted by the fourth silica gel column chromatographic separation is ethyl acetate-acetone solvent, the volume ratio of ethyl acetate to acetone in the ethyl acetate-acetone solvent is 1:2-1:3, and the elution mode is gradient elution; the eluent adopted by the chromatographic separation of the second SephadexLH-20 gel column is methanol; the mobile phase adopted by the semi-preparation high performance liquid chromatography separation is acetonitrile water solution with the acetonitrile volume fraction of 65%;
(5) Performing MCI (MCI-mediated color removal) on the component Fr.8.8 to obtain a component Fr.8.8.1;
sequentially performing fifth silica gel column chromatographic separation, third Sephadex LH-20 gel column chromatographic separation and fourth Sephadex LH-20 gel column chromatographic separation on the component Fr.8.8.1 to obtain a gastrodia elata active compound with a structure shown in a formula III; the eluent adopted by the fifth silica gel column chromatographic separation is chloroform-methanol solvent, the volume ratio of chloroform to methanol in the chloroform-methanol solvent is 6:1-0:1, and the elution mode is gradient elution; the eluent adopted by the third SephadexLH-20 gel column chromatography is a chloroform-methanol mixed solvent, and the volume ratio of chloroform to methanol in the chloroform-methanol mixed solvent is 1:1; the eluent adopted by the chromatographic separation of the fourth SephadexLH-20 gel column is methanol;
the step (4) and the step (5) have no time sequence.
In the present invention, all raw material components are commercially available products well known to those skilled in the art unless specified otherwise.
The invention carries out alcohol extraction on gastrodia elata to obtain alcohol extract; and extracting the ethanol extract with ethyl acetate, and concentrating to obtain an ethyl acetate phase. In the invention, the gastrodia elata is preferably a dried gastrodia elata flower stem. In the present invention, the alcohol extraction is preferably performed using an aqueous alcohol solution in which the volume fraction of alcohol is preferably 70 to 95%, more preferably 90 to 95%; the alcohol in the aqueous alcohol solution preferably comprises ethanol and/or methanol. In the invention, the ratio of the mass of the gastrodia elata to the volume of the alcohol-water solution is preferably 1kg:2 to 4L, more preferably 1kg:3L. In the present invention, the alcohol extraction is preferably reflux alcohol extraction; the number of the alcohol extractions is preferably 2 to 3, more preferably 3; the time of the single alcohol extraction is preferably 1 to 3 hours, more preferably 2 hours, and the extracting solutions are combined to obtain an alcohol extracting solution. In the present invention, the number of times of extraction of ethyl acetate is preferably 2 to 4 times, more preferably 3 times; the volume ratio of ethyl acetate to alcohol extract for single ethyl acetate extraction is preferably 1:0.5 to 2, more preferably 1:1 to 1.5. The concentration of the present invention is not particularly limited, and may be performed by any concentration means known to those skilled in the art, such as, for example, concentration under reduced pressure.
After obtaining an ethyl acetate phase, carrying out first silica gel column chromatographic separation on the ethyl acetate phase to obtain a component Fr.8; the eluent adopted by the first silica gel column chromatographic separation is petroleum ether-acetone solvent, the volume ratio of petroleum ether to acetone in the petroleum ether-acetone solvent is 100:1-0:1, and the elution mode is gradient elution. In a specific embodiment of the present invention, in the first silica gel column chromatographic separation process, TLC is preferably used to detect and combine the components with the same polarity (i.e. the fractions), so as to obtain 8 components, which are sequentially denoted as component fr.1, component fr.2, component fr.3, component fr.4, component fr.5, component fr.6, component fr.7 and component fr.8. In the present invention, the particle size of the first silica gel for chromatographic separation is preferably 200 to 300 mesh.
After the component Fr.8 is obtained, the component Fr.8 is subjected to second silica gel column chromatographic separation to obtain a component Fr.8.6 and a component Fr.8.8; the eluent adopted by the second silica gel column chromatographic separation is chloroform-acetone solvent, the volume ratio of chloroform to acetone in the chloroform-acetone solvent is 15:1-0:1, and the elution mode is gradient elution. In the present invention, the second silica gel column chromatography is preferably normal pressure normal phase silica gel column chromatography. In a specific embodiment of the present invention, the second silica gel column chromatographic separation process is preferably segmented to obtain 13 components, which are sequentially denoted as component fr.8.1, component fr.8.2, component fr.8.3, component fr.8.4, component fr.8.5, component fr.8.6, component fr.8.7, component fr.8.8, component fr.8.9, component fr.8.10, component fr.8.11, component fr.8.12 and component fr.8.13.
After the component Fr.8.6 is obtained, the invention carries out third silica gel column chromatography separation on the component Fr.8.6 to obtain the component Fr.8.6.6; the eluent adopted by the third silica gel column chromatographic separation is chloroform-acetone solvent, the volume ratio of chloroform to acetone in the chloroform-acetone solvent is 5:1-2:1, and the elution mode is gradient elution. In a specific embodiment of the present invention, the third silica gel column chromatographic separation process is preferably segmented to obtain 6 components, which are sequentially denoted as component fr.8.8.1, component fr.8.8.2, component fr.8.8.3, component fr.8.8.4, component fr.8.8.5 and component fr.8.8.6.
After the component Fr.8.6.6 is obtained, the invention sequentially carries out first Sephadex LH-20 gel column chromatography, fourth silica gel column chromatography, second Sephadex LH-20 gel column chromatography and semi-preparative high performance liquid chromatography on the component Fr.8.6.6 to obtain the gastrodia elata active compound with the structures shown in the formulas I and II; the eluent adopted by the first SephadexLH-20 gel column chromatography is a chloroform-methanol mixed solvent, and the volume ratio of chloroform to methanol in the chloroform-methanol mixed solvent is 1:1; the eluent adopted by the fourth silica gel column chromatographic separation is ethyl acetate-acetone solvent, the volume ratio of ethyl acetate to acetone in the ethyl acetate-acetone solvent is 1:2-1:3, and the elution mode is gradient elution; the eluent adopted by the chromatographic separation of the second SephadexLH-20 gel column is methanol; the semi-preparative High Performance Liquid Chromatography (HPLC) separation adopts a mobile phase of acetonitrile aqueous solution with the volume fraction of acetonitrile of 65 percent. In the present invention, the fourth silica gel column chromatography is preferably repeated normal pressure silica gel column chromatography, and the particle size of the silica gel used is preferably 200 to 300 mesh.
After the component Fr.8.8 is obtained, the component Fr.8.8 is subjected to MCI color removal to obtain the component Fr.8.8.1. In the invention, the eluent adopted for the MCI color removal is a water-methanol solvent, the volume ratio of water to methanol in the water-methanol solvent is 7:3-0:1, and the elution mode is preferably gradient elution. In a specific embodiment of the present invention, the MCI is preferably segmented during the color removal process to obtain 7 components, which are sequentially denoted as component fr.8.8.1, component fr.8.8.2, component fr.8.8.3, component fr.8.8.4, component fr.8.8.5, component fr.8.8.6 and component fr.8.8.7.
After the component Fr.8.8.1 is obtained, sequentially carrying out fifth silica gel column chromatography, third SephadexLH-20 gel column chromatography and fourth SephadexLH-20 gel column chromatography on the component Fr.8.8.1 to respectively obtain gastrodia elata active compound with a structure shown in a formula III; the eluent adopted by the fifth silica gel column chromatographic separation is chloroform-methanol solvent, and the volume ratio of chloroform to methanol in the chloroform-methanol solvent is 6:1-0:1; the eluent adopted by the third SephadexLH-20 gel column chromatography is a chloroform-methanol mixed solvent, and the volume ratio of chloroform to methanol in the chloroform-methanol mixed solvent is 1:1; the eluent adopted by the chromatographic separation of the fourth SephadexLH-20 gel column is methanol. In the present invention, the fifth silica gel column chromatography is preferably repeated normal pressure silica gel column chromatography, the elution mode is preferably gradient elution, and the particle size of the silica gel used is preferably 200 to 300 mesh.
The invention provides the gastrodia elata active compound disclosed by the technical scheme, the gastrodia elata active compound prepared by the preparation method disclosed by the technical scheme, or the application of any one of the gastrodia elata active compounds with structures shown in formulas IV-VII in preparation of medicines;
in the present invention, the use is preferably as cholinesterase inhibitor.
In the invention, the preparation method of the gastrodia elata active compound with the structure shown in the formula IV preferably comprises the following steps: and sequentially carrying out sixth silica gel column chromatographic separation and fifth SephadexLH-20 gel column chromatographic separation on the component Fr.8.8.2 to obtain the gastrodia elata active compound with the structure shown in the formula IV. In the present invention, the eluent used for the chromatographic separation of the fifth SephadexLH-20 gel column is preferably a chloroform-methanol mixed solvent, and the volume ratio of chloroform to methanol in the chloroform-methanol mixed solvent is preferably 1:1. In the invention, the eluent adopted by the sixth silica gel column chromatographic separation is preferably chloroform-methanol solvent, and the volume ratio of chloroform to methanol in the chloroform-methanol solvent is 15:1-1:1; the sixth silica gel column chromatography is preferably repeated normal pressure normal phase silica gel column chromatography, and the particle size of the adopted silica gel is preferably 200-300 meshes.
In the invention, the preparation method of the gastrodia elata active compound with the structure shown in the formula V preferably comprises the following steps: subjecting the component Fr.8.10 to seventh silica gel column chromatography to obtain a component Fr.8.10.10; and sequentially carrying out eighth silica gel column chromatographic separation, sixth Sephadex LH-20 gel column chromatographic separation and seventh Sephadex LH-20 gel column chromatographic separation on the component Fr.8.10.10 to obtain the gastrodia elata active compound with the structure shown in the formula V. In the invention, the eluent adopted by the seventh silica gel column chromatographic separation is preferably a chloroform-acetone mixed solvent, wherein the volume ratio of chloroform to acetone in the chloroform-acetone mixed solvent is 1:1; the seventh silica gel column chromatographic separation is preferably repeated normal pressure silica gel column chromatographic separation, and the particle size of the adopted silica gel is preferably 200-300 meshes, and in the specific embodiment of the invention, 12 components are preferably obtained by cutting, and are sequentially marked as a component Fr.8.10.1, a component Fr.8.10.2, a component Fr.8.10.3, a component Fr.8.10.4, a component Fr.8.10.5, a component Fr.8.10.6, a component Fr.8.10.7, a component Fr.8.10.8, a component Fr.8.10.9, a component Fr.8.10.10, a component Fr.8.10.11 and a component Fr.8.10.12. In the invention, the eluent adopted by the eighth silica gel column chromatographic separation is preferably chloroform-methanol solvent, the volume ratio of chloroform to methanol in the chloroform-methanol solvent is 15:1-1:1, and the elution mode is preferably gradient elution; the eighth silica gel column chromatography separation is preferably repeated normal pressure normal phase silica gel column chromatography, and the particle size of the adopted silica gel is preferably 200-300 meshes. In the present invention, the eluent used for the chromatographic separation of the sixth Sephadex LH-20 gel column is preferably chloroform-methanol solvent, and the volume ratio of chloroform to methanol in the chloroform-methanol solvent is preferably 1:1. In the present invention, the eluent used for the seventh Sephadex LH-20 gel column chromatographic separation is preferably methanol.
In the invention, the preparation method of the gastrodia elata active compound with the structures shown in the formulas VI and VII preferably comprises the following steps: subjecting the component Fr.7 to ninth silica gel column chromatography to obtain a component Fr.7.8 and a component Fr.7.11; sequentially performing tenth silica gel column chromatographic separation and semi-preparative HPLC separation on the component Fr.7.8 to obtain rhizoma Gastrodiae active compound with a structure shown in formula VII; and (3) performing eleventh silica gel column chromatographic separation and eighth SephadexLH-20 gel column chromatographic separation on the component Fr.7.11 to obtain the gastrodia elata active compound with the structure shown in the formula VI. In the invention, the eluent adopted by the ninth silica gel column chromatographic separation is preferably a petroleum ether-acetone mixed solvent, the volume ratio of petroleum ether and acetone in the petroleum ether-acetone mixed solvent is preferably 20:1-0:1, and the elution mode is preferably gradient elution; the ninth silica gel column chromatographic separation is preferably repeated normal pressure silica gel column chromatographic separation, and the particle size of the adopted silica gel is preferably 200-300 meshes, and in the specific embodiment of the invention, the ninth silica gel column chromatographic separation process is preferably segmented to obtain 14 components, which are sequentially marked as a component Fr.7.1, a component Fr.7.2, a component Fr.7.3, a component Fr.7.4, a component Fr.7.5, a component Fr.7.6, a component Fr.7.7, a component Fr.7.8, a component Fr.7.9, a component Fr.7.10, a component Fr.7.11, a component Fr.7.12, a component Fr.7.13 and a component Fr.7.14. In the invention, the eluent adopted by the tenth silica gel column chromatographic separation is preferably a petroleum ether-ethyl acetate solvent, the volume ratio of petroleum ether and ethyl acetate in the petroleum ether-ethyl acetate solvent is preferably 5:1-0:1, and the elution mode is preferably gradient elution; the tenth silica gel column chromatography separation is preferably repeated normal pressure normal phase silica gel column chromatography, and the particle size of the adopted silica gel is preferably 200-300 meshes. In the invention, the mobile phase adopted by the semi-preparative HPLC separation is a methanol aqueous solution with a methanol volume fraction of 75%. In the invention, the eluent adopted by the eleventh silica gel column chromatographic separation is preferably petroleum ether-ethyl acetate solvent, the volume ratio of petroleum ether and ethyl acetate in the petroleum ether-ethyl acetate solvent is preferably 20:1-0:1, and the elution mode is preferably gradient elution; the eleventh silica gel column chromatography is preferably repeated normal pressure normal phase silica gel column chromatography, and the particle size of the adopted silica gel is preferably 200-300 meshes. In the present invention, the eluent used for the eighth SephadexLH-20 gel column chromatography separation is preferably methanol.
The technical solutions of the present invention will be clearly and completely described in the following in connection with the embodiments of the present invention. It will be apparent that the described embodiments are only some, but not all, embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1
Reflux-extracting 170kg of dried gastrodia elata flower stems with 95% ethanol aqueous solution for 3 times, wherein each extraction time is 2h, combining 3 times of extracting solutions to obtain ethanol extracting solution, and the volume of ethanol aqueous solution for single extraction is 340L; the alcohol extract is extracted for 3 times by ethyl acetate in equal volume, and the ethyl acetate phase and the water phase are obtained by decompressing and concentrating the combined extract. And (3) performing silica gel column chromatography separation (the volume ratio of petroleum ether to acetone=20:1-0:1, and gradient elution) on the ethyl acetate phase, and combining parts with the same polarity (namely cutting) by TLC detection to obtain 8 components, wherein the 8 components are sequentially recorded as components Fr.1-Fr.8.
And (3) carrying out normal pressure normal phase silica gel column chromatography separation (petroleum ether: acetone volume ratio of (= 20:1-0:1) on the component Fr.7 (124 g), and segmenting to obtain 14 components which are sequentially recorded as components Fr.7.1-Fr.7.14. Component fr.7.8 (12 g) was separated by SephadexLH-20 gel column chromatography (chloroform: methanol volume ratio=1:1), followed by repeated normal pressure normal phase column chromatography (silica gel 200-300 mesh, chloroform: methanol volume ratio=15:1-0:1, gradient elution) and further semi-preparative HPLC separation (75% methanol aqueous solution isocratic elution) to give gastrodia elata active compound (5 mg) having the structure shown in formula VII. The component Fr.7.11 (20 g) is subjected to SephadexLH-20 gel column chromatography (chloroform: methanol volume ratio=1:1), and then repeated normal pressure normal phase column chromatography (silica gel 200-300 meshes, eluent is petroleum ether and ethyl acetate volume ratio=20:1-0:1) and SephadexLH-20 gel column chromatography (methanol) are carried out, so as to obtain the gastrodia elata active compound (22 mg) with the structure shown in the formula VI.
The component Fr.8 (743 g) was subjected to normal pressure normal phase silica gel column chromatography (chloroform: acetone volume ratio=15:1-0:1, gradient elution), and then divided into 13 fractions, which were sequentially designated as components Fr.8.1 to Fr.8.13. The component Fr.8.6 (26 g) is subjected to normal pressure normal phase silica gel column chromatography (chloroform: acetone volume ratio=5:1-2:1, gradient elution) and segmented to obtain 6 components, which are sequentially recorded as components Fr.8.6.1-Fr.8.6.6. Separating the component Fr.8.6.6 (5.3 g) by SephadexLH-20 gel column chromatography (chloroform: methanol volume ratio=1:1), repeated normal pressure normal phase column chromatography (silica gel 200-300 meshes, ethyl acetate: acetone volume ratio=1:2-1:3, gradient elution) and SephadexLH-20 gel column chromatography separation (methanol), further semi-preparative HPLC separation (isocratic elution of 65v/v% acetonitrile water solution for 15min, isocratic elution of 70% acetonitrile water solution for 20 min), and obtaining the gastrodia elata active compound with the structures shown in the formula I (23 mg) and the formula II (4 mg). The component fr.8.8 (13 g) was subjected to MCI-based color removal (water: methanol volume ratio=7:3-0:1, gradient elution) to obtain 7 components, which were sequentially noted as component fr.8.8.1 to component fr.8.8.7. The fraction Fr.8.8.1 was subjected to repeated normal pressure normal phase column chromatography (silica gel 200-300 mesh, chloroform: methanol volume ratio=6:1-0:1, gradient elution), sephadexLH-20 gel column chromatography (chloroform: methanol volume ratio=1:1) and SephadexLH-20 gel column chromatography (methanol) to obtain a gastrodia elata active compound (29 mg) having the structure shown in formula III. The component Fr.8.8.2 is subjected to repeated normal pressure normal phase column chromatography separation (silica gel 200-300 meshes, chloroform: methanol volume ratio=15:1-1:1, gradient elution), sephadexLH-20 gel column chromatography (chloroform: methanol volume ratio=1:1) and SephadexLH-20 gel column chromatography (methanol) to obtain a gastrodia elata active compound (53 mg) with a structure shown in a formula IV. And (3) carrying out normal pressure normal phase silica gel column chromatography separation (chloroform: acetone volume ratio=1:1, silica gel 200-300 meshes) on the component Fr.8.10, and segmenting to obtain 12 parts, which are sequentially recorded as components Fr.8.10.1-Fr.8.10.12. Subjecting the component Fr.8.10.10 to normal pressure normal phase column chromatography (silica gel 200-300 meshes, volume ratio of chloroform to methanol=15:1-1:1, gradient elution), sephadex LH-20 gel column chromatography (chloroform: methanol volume ratio=1:1) and Sephadex LH-20 gel column chromatography (methanol) to obtain rhizoma Gastrodiae active compound (16 mg) with the structure shown in formula V.
Of the gastrodia elata active compounds having the structures shown in formulas I to VII, the gastrodia elata active compounds having the structures shown in formulas I (gastrodinolB) and II (gastrodinolC) are gibberellin compounds, the gastrodia elata active compounds having the structures shown in formulas III (benzylmulberrofuranG), IV (mulberrofuranG) and V (mulberrofuran) are Phellin compounds, and the gastrodia elata active compounds having the structures shown in formulas VI (morusin) and VII (notabilisinG) are isopentenyl flavonoid compounds, and the nuclear magnetic data of each compound are as follows:
GastrodinolB (formula I): c (C) 35 H 28 O 7 ；(-)HRESI-MSm/z673.1695([M+CF 3 COO] - ,calcd673.1691)； 1 HNMR(500MHz,DMSO-d 6 )δ H 12.11(s,1H,10-OH),9.12(s,3H,4',4”,4”'-OH),8.21(d,J＝9.0Hz,1H,H-7),7.50(d,J＝8.9Hz,1H,H-6),7.02(s,1H,H-11),6.80(d,J＝8.6Hz,2H,H-2',H-6'),6.71(d,J＝8.5Hz,4H,H-2”,H-6”,H-2”',H-6”'),6.55(d,J＝8.5Hz,2H,H-3',H-5'),6.38(d,J＝8.5Hz,4H,H-3”,H-5”,H-3”',H-5”'),4.10(s,2H,H-7'),3.92(s,3H,12-OCH 3 ),3.14(m,4H,H-7”,H-7”')； 13 CNMR(126MHz,DMSO)δ C 210.38(C-3),200.57(C-1),159.72(C-12),155.98(C-4”,C-4”'),155.40(C-4'),155.32(C-10),142.19(C-5),140.34(C-4),136.80(C-8),135.46(C-7),130.49(C-1'),130.44(C-2”,C-6”,C-2”',C-6”'),128.72(C-2',C-6'),125.15(C-1”,C-1”'),117.65(C-6),115.16(C-3”,C-5”,C-3”',C-5”'),115.06(C-13),114.97(C-3',C-5'),112.56(C-9),101.84(C-11),61.84(C-2),56.35(12-OCH 3 ),39.96(C-7”,C-7”'),28.73(C-7')。
Gastrodinol C (formula II) C 35 H 28 O 7 ；(-)HRESI-MSm/z605.1812([M+COOH] - ,calcd605.1817)； 1 HNMR(500MHz,Methanol-d 4 )δ H 7.93(dt,J＝8.5,1.6Hz,1H,H-7),7.43(d,J＝8.4Hz,1H,H-6),7.09(d,J＝8.6Hz,2H,H-2',H-6'),6.89(s,1H,H-13),6.80(d,J＝8.6Hz,4H,H-2”,H-6”,H-2”',H-6”'),6.65(d,J＝8.5Hz,2H,H-3',H-5'),6.38(d,J＝8.6Hz,4H,H-3”,H-5”,H-3”',H-5”'),4.05(s,2H,H-7'),3.89(s,3H,12-OCH 3 ),3.21(s,4H,H-7”,H-7”')； 13 CNMR(126MHz,MeOD)δ C 211.92(C-3),203.22(C-1),162.13(C-12),157.27(C-4”,C-4”'),156.20(C-4'),154.56(C-10),144.67(C-5),141.66(C-4),139.67(C-8),139.45(C-7),133.10(C-1'),131.81(C-2”,C-6”,C-2”',C-6”'),130.60(C-2',C-6'),127.45(C-1”,C-1”'),120.59(C-11),118.03(C-6),115.89(C-3”,C-5”,C-3”',C-5”'),115.80(C-3',C-5'),115.37(C-9),100.75(C-13),63.89(C-2),56.31(12-OCH 3 ),41.76(C-7”,C-7”'),29.32(C-7')。
Benzylmullorofurang (formula III) C 41 H 32 O 9 ；(+)HRESI-MSm/z669.2126([M+H] + ,calcd669.2119)； 1 HNMR(500MHz,Acetone-d 6 )δ H 8.56(s,1H,10”-OH),8.54(s,1H,6-OH),8.50(s,1H,12”-OH),8.34(s,1H,18”-OH),8.12(s,1H,4”'-OH),7.49(s,1H,3'-OH),7.33(d,J＝8.4Hz,1H,H-4),7.23(d,J＝8.7Hz,1H,H-14”),7.14(dd,J＝8.4,1.2Hz,1H,H-20”),7.03(s,1H,H-6'),6.94(dd,J＝2.1,0.9Hz,1H,H-7),6.90(d,J＝8.6Hz,2H,H-2”',H-6”'),6.77(dd,J＝8.4,2.1Hz,1H,H-5),6.67(d,J＝8.6Hz,2H,H-3”',H-5”'),6.65(d,J＝0.9Hz,1H,H-3),6.50(dd,J＝8.4,2.5Hz,1H,H-19”),6.43(dd,J＝3.7,2.4Hz,1H,H-2”),6.42(d,J＝2.4Hz,1H,H-11”),6.36(d,J＝2.5Hz,1H,H-17”),6.24(dd,J＝8.6,2.4Hz,1H,H-13”),4.16(s,2H,H-7”'),3.56(d,J＝5.6Hz,1H,H-3”),3.36(dd,J＝11.9,5.6Hz,1H,H-4”),2.99(m,1H,H-5”),2.73(dd,J＝17.1,5.4Hz,1H,H-6”a),2.02(dd,J＝17.1,12.0Hz,1H,H-6”b),1.77(s,3H,H-7”). 13 CNMR(126MHz,Acetone)δ C 159.93(C-12”),157.78(C-18”),157.59(C-10”),156.64(C-6),156.45(C-7a),156.33(C-4”'),155.78(C-3'),154.60(C-2),153.38(C-16”),152.35(C-5'),134.39(C-1”),131.98(C-1”'),131.49(C-1'),130.47(C-14”),129.83(C-2”',C-6”'),127.91(C-20”),122.38(C-3a),122.34(C-2”),121.99(C-4),119.70(C-2'),117.56(C-15”),116.89(C-9”),115.94(C-3”',C-5”'),115.28(C-4'),113.09(C-5),109.86(C-19”),109.56(C-6'),107.15(C-13”),105.60(C-3),104.62(C-11”),103.95(C-17”),102.47(C-8”),98.28(C-7),37.40(C-4”),36.29(C-6”),35.16(C-3”),32.05(C-7”'),28.52(C-5”),23.96(C-7”)。
mulberrofuranG (formula IV): 600MHz, Methanol-d 4 δ H :7.11(d,J＝8.2Hz,1H,H-6'),6.60(d,J＝9.9Hz,1H,H-1”),6.42(d,J＝2.2Hz,1H,H-3'),6.41–6.39(dd,J＝8.2Hz,2.2Hz,1H,H-5'),6.15(s,1H,H-6),5.59(d,J＝9.9Hz,1H,H-2”),5.09(t,J＝7.1Hz,1H,H-2”'),3.10(d,J＝7.1Hz,2H,H-1”'),1.59(s,3H,H-4”'),1.43(s,6H,H-4”,H-5”),1.40(s,3H,H-5”'). 13 CNMR(151MHz,MeOD)δ C :183.88(C-4),163.58(C-2),162.72(C-5),162.07(C-4'),160.48(C-7),157.95(C-2'),153.79(C-8a),132.87(C-3”'),132.44(C-6'),128.20(C-2”),122.72(C-2”'),122.05(C-3),115.77(C-1”),113.07(C-1'),108.04(C-5'),105.92(C-8),103.84(C-3'),102.22(C-4a),100.11(C-6),79.14(C-3”),28.40(C-4”,C-5”),25.86(C-4”'),24.90(C-1”'),17.68(C-5”')。
mulberrofuran (formula V): 500MHz, methanol-d 4 δ H :7.11(d,J＝8.3Hz,1H,H-6'),6.62(d,J＝10.0Hz,1H,H-1”),6.38(d,J＝8.3Hz,1H,H-5'),6.37(s,1H,H-3'),6.16(s,1H,H-6),6.03(s,1H,H-4”'b),5.79(s,1H,H-4”'a),5.61(d,J＝10.0Hz,1H,H-2”),3.82(s,2H,H-1”'),1.83(s,3H,H-5”'),1.44(s,6H,H-4”,H-5”)； 13 CNMR(126MHz,MeOD)δ C :200.68(C-2”'),183.28(C-4),164.32(C-2),162.61(C-5),162.51(C-2'),160.69(C-7),158.06(C-4'),153.84(C-8a),145.54(C-3”'),132.32(C-6'),128.43(C-2”),125.71(C-4”'),117.38(C-3),115.69(C-1”),112.31(C-1’),108.38(C-5'),105.54(C-4a),103.84(C-3'),102.38(C-8),100.33(C-6),79.23(C-3”),35.73(C-1”'),28.39(C-4”,C-5”),17.83(C-5”')。
Morusin (formula VI): 500MHz, acetone-d 6 δ H :8.55(d,J＝9.0Hz,1H,H-14”),7.34(d,J＝8.4Hz,1H,H-4),6.97(d,J＝8.4Hz,1H,H-20”),6.93(d,J＝2.2Hz,1H,H-7),6.92(d,J＝0.9Hz,1H,H-3),6.77(s,2H,H-2',H-6'),6.76(dd,J＝8.3,2.1Hz,1H,H-5),6.52(d,J＝2.4Hz,1H,H-17”),6.37(dd,J＝9.0,2.4Hz,1H,H-13”),6.30(dd,J＝8.4,2.4Hz,1H,H-19”),6.23(d,J＝2.3Hz,1H,H-11”),5.76(d,J＝2.8Hz,1H,H-2”),4.62(dd,J＝5.4,3.7Hz,1H,H-4”),4.12(p,J＝2.6Hz,1H,H-3”),3.75(dt,J＝6.3,3.0Hz,1H,H-5”),2.53-2.43(m,1H,H-6”b),2.26-2.14(m,1H,H-6”a),1.95(s,3H,H-7”)； 13 CNMR(126MHz,Acetone)δ C :209.36(C-8”),167.01(C-12”),166.73(C-10”),157.91(C-16”),157.80(C-18”),156.63(C-3',5',2),156.49(C-7a),155.36(C-6),135.07(C-14”),133.71(C-1”),130.83(C-1'),128.60(C-20”),124.38(C-2”),122.45(C-15”),121.78(C-4),121.74(C-3a),116.48(C-9”),113.23(C-4'),113.09(C-5),109.05(C-13”),107.29(C-19”),104.69(C-2',C-6'),103.55(C-11”),103.48(C-17”),101.80(C-3),98.30(C-7),47.77(C-4”),36.40(C-5”),33.08(C-3”),32.27(C-6”),23.82(C-7”)。
notabilisineg (formula VII): 400MHz, methanol-d 4 δ H :7.34(d,J＝8.4Hz,1H,H-4),7.14(d,J＝8.6Hz,1H,H-14”),7.09(d,J＝8.4Hz,1H,H-20”),6.92(s,1H,H-7),6.91(d,J＝1.8Hz,2H,H-2',H-6'),6.82(s,1H,H-3),6.73(dd,J＝8.4,2.1Hz,1H,H-5),6.46(dd,J＝8.4,2.6Hz,1H,H-19”),6.40(d,J＝4.7Hz,1H,H-2”),6.35(d,J＝2.5Hz,1H,H-17”),6.33(d,J＝2.4Hz,1H,H-11”),6.14(dd,J＝8.6,2.4Hz,1H,H-13”),3.34–3.23(m,2H,H-3”,H-4”),2.95(td,J＝11.1,5.4Hz,1H,H-5”),2.66(dd,J＝17.1,5.5Hz,1H,H-6”a),2.01(dd,J＝17.0,11.4Hz,1H,H-6”b),1.78(s,3H,H-7”)； 13 CNMR(101MHz,MeOD)δ C :158.65(C-10”),157.02(C-18”),156.51(C-12”),156.33(C-6),155.85(C-2),155.39(C-5'),154.34(C-3'),153.52(C-16”),152.21(C-7a),132.52(C-1”),130.14(C-1'),129.18(C-14”),126.60(C-20”),121.96(C-2”),121.71(C-3a),120.60(C-4),116.89(C-4'),115.96(C-15”),112.52(C-9”),111.85(C-5),108.65(C-19”),105.63(C-13”),104.10(C-3),103.60(C-6'),103.15(C-2'),102.78(C-17”),101.70(C-8”),100.76(C-11”),97.10(C-7),36.24(C-3”),35.34(C-6”),34.02(C-5”),27.44(C-4”),22.54(C-7”)。
Test example 1
Acetylcholinesterase inhibitory activity of gastrodia elata active compound having structure shown in formulas I-VII
(1) Experimental materials and reagents
Reagent: na (Na) 2 HPO 4 ；NaH 2 PO 4 The method comprises the steps of carrying out a first treatment on the surface of the Acetylcholinesterase, thiocholine iodide, DTNB, tacrine (Tacrine) were all purchased from Sigma.
Instrument: multiskanFC microplate reader (Thermo Corp.).
(2) Experimental method
(2.1) phosphate buffer (0.1 MNA per 100mL of phosphate buffer) 2 HPO 4 94.7mL of solution; 0.1MNaH 2 PO 4 5.3mL of solution, adjusting the pH value to 8.0) to dilute AChE into 0.1U/mL of working solution;
(2.2) preparation of a solution (working solution) of 6.25mM of the thioacetylcholine iodide and DTNB in phosphate buffer;
(2.3) Compound diluted with DMSO to a concentration gradient. The positive control is tacrine, diluted with DMSO to a concentration gradient; the negative control group (NC group) was a 2% dmso solvent control.
(2.4) reactions were performed in 96-well plates, plated at 200. Mu.L/system, 3 replicates per sample;
(2.5) setting of concentration gradients of Compound and Tacrine (TA) (diluted with 1% DMSO) As shown in Table 1, individual samples with better activity were reduced in concentration to determine IC 50 。
Setting of the concentration gradients of the compounds of Table 1 and Tacrine (TA)
(2.6) plating: 200. Mu.L/system, final DMSO concentration in each well was 0.1%, 3 duplicate wells were made per sample, and the order and amounts of reagents added are shown in Table 2.
TABLE 2 order and amount of addition of the respective reagents
(2.7) absorbance at 405nm was measured every 30 seconds within 1 hour after addition of the developer and substrate.
(2.8) selecting a sample absorbance at an NC group absorbance average of about 1, calculating a compound absorbance average (compound measurement value-background value), and calculating a compound AChE inhibition ratio as (NC-compound absorbance average)/NC x 100%.
(3) The experimental results are shown in table 3.
TABLE 3 results of acetylcholinesterase inhibitory Activity of Gastrodia elata active Compounds
/>
/>
As shown in Table 3, the gastrodia elata active compound prepared by the invention has good inhibition effect on acetylcholinesterase.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (8)
1. A compound having the structure of formulae I to III, formula V:
2. a process for the preparation of a compound as claimed in claim 1, comprising the steps of:
(1) Extracting gastrodia elata with alcohol to obtain an alcohol extract; extracting the ethanol extract with ethyl acetate, and concentrating to obtain an ethyl acetate phase;
(2) Subjecting the ethyl acetate phase to first silica gel column chromatography separation to obtain a component Fr.8; the eluent adopted by the first silica gel column chromatographic separation is petroleum ether-acetone solvent, the volume ratio of petroleum ether to acetone in the petroleum ether-acetone solvent is 100:1-0:1, and the elution mode is gradient elution;
(3) Subjecting the component Fr.8 to a second silica gel column chromatography to obtain 13 components, which are sequentially named as a component Fr.8.1, a component Fr.8.2, a component Fr.8.3, a component Fr.8.4, a component Fr.8.5, a component Fr.8.6, a component Fr.8.7, a component Fr.8.8, a component Fr.8.9, a component Fr.8.10, a component Fr.8.11, a component Fr.8.12 and a component Fr.8.13; the eluent adopted by the second silica gel column chromatographic separation is chloroform-acetone solvent, the volume ratio of chloroform to acetone in the chloroform-acetone solvent is 15:1-0:1, and the elution mode is gradient elution;
(4) Subjecting the component Fr.8.6 to third silica gel column chromatography to obtain a component Fr.8.6.6; the eluent adopted by the third silica gel column chromatographic separation is chloroform-acetone solvent, the volume ratio of chloroform to acetone in the chloroform-acetone solvent is 5:1-2:1, and the elution mode is gradient elution;
sequentially carrying out first SephadexLH-20 gel column chromatography, fourth silica gel column chromatography, second SephadexLH-20 gel column chromatography and semi-preparative high performance liquid chromatography on the component Fr.8.6.6 to respectively obtain compounds with structures shown in a formula I and a formula II; the eluent adopted by the first SephadexLH-20 gel column chromatography is a chloroform-methanol mixed solvent, and the volume ratio of chloroform to methanol in the chloroform-methanol mixed solvent is 1:1; the eluent adopted by the fourth silica gel column chromatographic separation is ethyl acetate-acetone solvent, the volume ratio of ethyl acetate to acetone in the ethyl acetate-acetone solvent is 1:2-1:3, and the elution mode is gradient elution; the eluent adopted by the chromatographic separation of the second SephadexLH-20 gel column is methanol; the mobile phase adopted by the semi-preparation high performance liquid chromatography separation is acetonitrile water solution with the acetonitrile volume fraction of 65%;
(5) Performing MCI (MCI-mediated color removal) on the component Fr.8.8 to obtain a component Fr.8.8.1;
sequentially carrying out fifth silica gel column chromatographic separation, third SephadexLH-20 gel column chromatographic separation and fourth SephadexLH-20 gel column chromatographic separation on the component Fr.8.8.1 to obtain a compound with a structure shown in a formula III; the eluent adopted by the fifth silica gel column chromatographic separation is chloroform-methanol solvent, the volume ratio of chloroform to methanol in the chloroform-methanol solvent is 6:1-0:1, and the elution mode is gradient elution; the eluent adopted by the third SephadexLH-20 gel column chromatography is a chloroform-methanol mixed solvent, and the volume ratio of chloroform to methanol in the chloroform-methanol mixed solvent is 1:1; the eluent adopted by the chromatographic separation of the fourth SephadexLH-20 gel column is methanol;
(6) Subjecting the component Fr.8.10 to seventh silica gel column chromatography to obtain a component Fr.8.10.10; sequentially carrying out eighth silica gel column chromatographic separation, sixth SephadexLH-20 gel column chromatographic separation and seventh SephadexLH-20 gel column chromatographic separation on the component Fr.8.10.10 to obtain a compound with a structure shown in a formula V; the eluent adopted by the seventh silica gel column chromatographic separation is a chloroform-acetone mixed solvent, and the volume ratio of chloroform to acetone in the chloroform-acetone mixed solvent is 1:1; the eluent adopted by the eighth silica gel column chromatographic separation is chloroform-methanol solvent, the volume ratio of chloroform to methanol in the chloroform-methanol solvent is 15:1-1:1, and the elution mode is gradient elution; the eluent adopted by the chromatographic separation of the sixth SephadexLH-20 gel column is chloroform-methanol solvent, and the volume ratio of chloroform to methanol in the chloroform-methanol solvent is 1:1; the eluent adopted by the seventh SephadexLH-20 gel column chromatographic separation is methanol;
the step (4) and the step (5) have no time sequence.
3. The method according to claim 2, wherein the alcohol extraction is performed with an aqueous alcohol solution; the alcohol in the alcohol-water solution comprises ethanol and/or methanol; the volume fraction of the alcohol in the alcohol water solution is 70-95%.
4. The preparation method according to claim 3, wherein the ratio of the mass of the gastrodia elata to the volume of the aqueous alcohol solution is 1kg: 2-4L.
5. The process according to claim 2, 3 or 4, wherein the alcohol extraction is reflux alcohol extraction; the times of the alcohol extraction are 2-3 times, and the time of the single alcohol extraction is 1-3 hours.
6. The preparation method according to claim 2, wherein the eluent used for MCI color removal is a water-methanol solvent, and the volume ratio of water to methanol in the water-methanol solvent is 7:3-0:1.
7. Use of a compound of formula I-III as defined in claim 1 or formula V as defined in any one of claims 2-6 for the preparation of a cholinesterase inhibitor.
8. The use according to claim 7, wherein the use is anti-senile dementia.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211104188.5A CN115304616B (en) | 2022-09-09 | 2022-09-09 | Gastrodia elata active compound and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211104188.5A CN115304616B (en) | 2022-09-09 | 2022-09-09 | Gastrodia elata active compound and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115304616A CN115304616A (en) | 2022-11-08 |
| CN115304616B true CN115304616B (en) | 2023-08-04 |
Family
ID=83866217
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211104188.5A Active CN115304616B (en) | 2022-09-09 | 2022-09-09 | Gastrodia elata active compound and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115304616B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115286606A (en) * | 2022-09-09 | 2022-11-04 | 中国科学院昆明植物研究所 | Gastrodia elata flavonoid compound and preparation method and application thereof |
-
2022
- 2022-09-09 CN CN202211104188.5A patent/CN115304616B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115286606A (en) * | 2022-09-09 | 2022-11-04 | 中国科学院昆明植物研究所 | Gastrodia elata flavonoid compound and preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| Potential Cancer- and Alzheimer’s Disease-Targeting Phosphodiesterase Inhibitors from Uvaria alba: Insights from In Vitro and Consensus Virtual Screening;Mark Tristan Quimque等;ACS Omega;第6卷;8403−8417 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115304616A (en) | 2022-11-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN115073295B (en) | Unsaturated fatty acid in purslane, and extraction and separation method and application thereof | |
| CN109879844B (en) | Method for extracting and separating seven flavonoid chemical components in caulis Sinomenii Microphylli | |
| CN102210726A (en) | Method for combined extraction of mallow flavones and mallow saponins in seashore mallow and application | |
| CN108084060B (en) | Alkaloid oleraurea in purslane and extraction and separation method thereof | |
| CN111704544B (en) | Labdane diterpenoid compound and separation method and application thereof | |
| CN113754533A (en) | Oxidized labdane diterpenoid compounds and separation method and application thereof | |
| CN103169727B (en) | General-flavonoid compound in chionanthus as well as preparation method and application thereof | |
| CN108689851B (en) | Tiglic alkane type diterpene compound and preparation method and application thereof | |
| Wang et al. | Purine-containing cucurbitane triterpenoids from Cucurbita pepo cv dayangua | |
| CN115286606B (en) | Gastrodia elata flavonoid compound and preparation method and application thereof | |
| CN115304616B (en) | Gastrodia elata active compound and preparation method and application thereof | |
| Thomas et al. | Alstonoside, a secoiridoid glucoside from Alstonia scholaris | |
| Shul'ts et al. | Flavonoids of roots of Glycyrrhiza uralensis growing in Siberia | |
| CN106565444B (en) | The extracting method and application of Chinese yam aerial part phenanthrene class compound | |
| CN103113439A (en) | Method for preparing kaempferol-3-O-Beta-D-glucuronide in euphorbia sororia | |
| CN109265503B (en) | Flavonoid compound and preparation method and application thereof | |
| CN103665090B (en) | Ilex cornuta saponin compound, its preparation method and application | |
| CN116333029B (en) | Tea seed saponin compound and application thereof | |
| CN111548255A (en) | Diterpenoid compounds in taxus chinensis, preparation method thereof and application thereof in pharmacy | |
| CN112194690B (en) | 3 compounds in radix Rubiae and extraction and separation method | |
| CN110251552B (en) | Preparation method of total flavonoids of sauropus spatulifolius | |
| CN114920796B (en) | Steroid saponin compound extracted from paris polyphylla, preparation and application | |
| CN114276339B (en) | Sesquilignans compound, and separation method and application thereof | |
| CN114702535B (en) | Active ingredient in Potentilla mucida and preparation method and application thereof | |
| CN114177164B (en) | Preparation method and application of patrinia villosa anthracenol I |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| OL01 | Intention to license declared | ||
| OL01 | Intention to license declared |